The let-99 gene is required for proper spindle orientation during cleavage of the C. elegans embryo.
The orientation of cell division is a critical aspect of development. In 2-cell C. elegans embryos, the spindle in the posterior cell is aligned along the long axis of the embryo and contributes to the unequal partitioning of cytoplasm, while the spindle in the anterior cell is oriented transverse to the long axis. Differing spindle alignments arise from blastomere-specific rotations of the nuclear-centrosome complex at prophase. We have found that mutations in the maternally expressed gene let-99 affect spindle orientation in all cells during the first three cleavages. During these divisions, the nuclear-centrosome complex appears unstable in position. In addition, in almost half of the mutant embryos, there are reversals of the normal pattern of spindle orientations at second cleavage: the spindle of the anterior cell is aligned with the long axis of the embryo and nuclear rotation fails in the posterior cell causing the spindle to form transverse to the long axis. In most of the remaining embryos, spindles in both cells are transverse at second cleavage. The distributions of several asymmetrically localized proteins, including P granules and PAR-3, are normal in early let-99 embryos, but are perturbed by the abnormal cell division orientations at second cleavage. The accumulation of actin and actin capping protein, which marks the site involved in nuclear rotation in 2-cell wild-type embryos, is abnormal but is not reversed in let-99 mutant embryos. Based on these data, we conclude that let-99(+) is required for the proper orientation of spindles after the establishment of polarity, and we postulate that let-99(+) plays a role in interactions between the astral microtubules and the cortical cytoskeleton.
Rose
L S
LS
Section of Genetics and Development, Cornell University, Ithaca, NY 14853, USA. lsrose@ucdavis.edu
Kemphues
K
K
D000199:Actins / Q000378:metabolism; D000818:Animals; D017173:Caenorhabditis elegans / Q000166:cytology / Q000196:embryology*; D029742:Caenorhabditis elegans Proteins; D002455:Cell Division / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D015801:Helminth Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D007150:Immunohistochemistry; D008858:Microscopy, Phase-Contrast; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D017346:Protein Serine-Threonine Kinases; D008941:Spindle Apparatus / Q000235:genetics* / Q000648:ultrastructure
D000199:Actins; D029742:Caenorhabditis elegans Proteins; D015801:Helminth Proteins; C087977:par-2 protein, C elegans; C097238:PAR-3 protein, C elegans; D017346:Protein Serine-Threonine Kinases